Comparative Proteomics Analysis of Pig Muscle Exudate through Label-Free Liquid Chromatography-Mass Spectrometry

Abstract

Capital-driven animal husbandry systems undertaken in the last century led to the abandoning of many pig breeds that were not profitable. These local pig breeds and their respective production systems have great potential as they are able to respond to the high criteria and needs of modern society concerning some environmental aspects, animal-welfare, healthiness, etc. This is the case of the black pigs of Italy. The Apulo-Calabrese is a breed of black pig, known by many other names such as Nero d’Abruzzo. In order to further understand the biological differences between different types of porcine genetics (Nero d’Abruzzo and commercial-hybrid) we used a label-free LC-MS strategy and Western-blot to characterize the proteomes of muscle-exudate collected from these pigs. This proteomics approach identified 1669 proteins of which 100 changed significantly in abundance between breeds. Bioinformatics functional analysis indicated that differentially expressed proteins were involved in several biological processes related to energy-metabolism and response to oxidative stress, suggesting that these functions might distinguish between these pigs. Fatty-acid synthase, catalase and glutathione-peroxidase, involved in enzymatic activity were found to be more represented in samples obtained from the Nero d’Abruzzo. This biological information can potentially provide new biological factors that could determine the different production performances of these pigs, distinguished by their different genetic backgrounds.

Label-Free Quantitative Analysis of Pig Liver Proteome after Hepatitis E Virus Infection

“…Liver exudate was obtained following the protocol previously used for other tissues [ 17 ]. Briefly, three cubes of about 10 g each were taken from the liver sampled for each animal.…”

“…The applied gradient initially envisaged the use of 98% of A and 2% of B; subsequently, the percentage of B was increased up to 32% in a time interval of 75 min, and then further increased up to 90% in 5 min; this condition then remained stable for a further 5 min with a flow rate of 300 nL/min. The acquisition of MS1 and MS2 spectra was performed as recently described by Di Luca et al [ 17 ].…”